1. Field of the Invention
The present invention relates generally to the field of chemistry and molecular biology. More particularly, it concerns emulsion formulations and methods for emulsion amplification of nucleic acid.
2. Description of Related Art
Emulsion PCR allows for the parallel analysis of millions of compartmentalized polynucleotide molecules within a single reaction vessel. BEAMing (beads, emulsions, amplification, and magnetics) built on emulsion PCR by keeping products formed within each compartment together once the emulsions are broken. This was accomplished through (i) inclusion of beads within the compartments and (ii) ensuring that at least one strand of the PCR product is bound to the beads. After amplification, each bead is coated with thousands of copies of the single DNA molecule present in the compartment that contained the bead and, after emulsion breaking, these beads could easily be recovered with a magnet or by centrifugation.
Beads obtained via BEAMing accurately reflect the polynucleotide diversity present in the template population and can be used to determine what fraction of a polynucleotide population contains a specific sequence variation. Because each bead contains thousands of molecules of the identical sequence, the signal to noise ratio obtained with methods to determine the polynucleotide sequence is extremely high. Millions of beads can be analyzed within minutes using conventional flow cytometry or sequencing instruments. However, conventional BEAMing techniques remain cumbersome and have not been successfully adapted for high throughput analyses. Thus, there remains a need for new emulsion reagents and techniques to address these issues.